ATCC medium: 2636 Thermalkalibacillus uzonensis Medium
NH4Cl ....................................1.00 g
NaCl.....................................3.60 g
KCl......................................1.00 g
Na2SO4 ...................................0.50 g
CaCl2 ....................................0.04 g
MgCl2 . 6H2O .............................0.04 g
KH2PO4 ...................................0.50 g
K2HPO4 ...................................1.00 g
Wolfe's Mineral Solution (see below).....1.00 ml
Wolfe's Vitamin Solution (see below).....1.00 ml
Yeast extract............................0.50 g
Distilled water..........................1.0 L
The pH of the medium should be adjusted to 8.3. Autoclave at 121?C for 15 minutes. Filter sterilize and
prepare separately a 2M pyruvate stock solution. Add 100ul of the pyruvate stock solution for every 10ml of
media.
Wolfe's Mineral Solution:
Available from ATCC as a sterile ready-to-use liquid (Trace Mineral Supplement, catalog no. MD-TMS.)
Nitrilotriacetic acid........1.500 g
MgSO4 . 7H2O .................3.000 g
MnSO4 . H2O ..................0.500 g
NaCl.........................1.000 g
FeSO4 . 7H2O .................0.100 g
Co(NO3)2 . 6H2O...............0.100 g
CaCl2 ........................0.100 g
ZnSO4 . 7H2O .................0.100 g
CuSO4 . 5H2O .................0.010 g
AlK(SO4)2 . 12H2O.............0.010 g
H3BO3 ........................0.010 g
Na2MoO4 . 2H2O................0.010 g
Na2SeO3 (anhydrous) ..........0.001 g
Na2WO4. 2H2O..................0.010 g
NiCl2 . 6H2O .................0.020 g
Distilled water..............1.0 L Add nitrilotriacetic acid to approximately 500 ml of water and adjust to pH 6.5 with KOH to dissolve the
compound. Bring volume to 1.0 L with remaining water and add remaining compounds one at a time.
Wolfe's Vitamin Solution:
Available from ATCC as a sterile ready-to-use liquid (Vitamin Supplement, catalog no. MD-VS).
Biotin.............................2.0 mg
Folic acid.........................2.0 mg
Pyridoxine hydrochloride..........10.0 mg
Thiamine . HCl.....................5.0 mg
Riboflavin.........................5.0 mg
Nicotinic acid.....................5.0 mg
Calcium D-(+)-pantothenate.........5.0 mg
Vitamin B12........................0.1 mg
p-Aminobenzoic acid................5.0 mg
Thioctic acid......................5.0 mg
Distilled water....................1.0 L